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Biochemical and immunological properties of the human carcinoma-associated CAR-3 epitope defined by the monoclonal antibody AR-3.

Abstract
The monoclonal antibody AR-3 reacts with an epitope (CAR-3) carried on a high-molecular-weight glycoprotein associated with carcinomas of the pancreas, stomach, colon, uterus, and ovary. This study reports the partial purification and characterization of CAR-3-bearing molecule. The antigen was quantified by a double determinant immunoradiometric assay. CAR-3 antigen was purified by a three-step procedure, consisting of perchloric acid extraction, molecular sieving on Sepharose CL-4B, and affinity chromatography on AR-3 antibodies coupled to Sepharose 4B. Following this procedure CAR-3 antigen was purified about 400-fold with a 36% yield. Treatment of the CAR-3 antigen with 16 mM metaperiodate or with 1 N NaOH resulted in complete loss of activity. Antigenicity survived enzymatic treatments known to destroy proteins. The epitope was found to be carried on a molecule with a molecular weight of greater than 400,000 with a density of 1.45 g/ml, metabolically labeled with [35S]sulfate, [3H]glucosamine, and [35S]methionine. It is concluded that CAR-3 epitope is expressed on a carbohydrate moiety linked to a sulfo-mucin-like molecule via an O-glycosidic bond. Cross-competition experiments showed that CAR-3 epitope is strictly related or in close topographic proximity to Lewis(a) and Lewis(b) antigens. Cross-double determinant immunoradiometric assay experiments indicated that the same mucin carrying CAR-3 bears also CA 19-9, CA 125, and BW 494 epitopes.
AuthorsM Prat, E Medico, P Rossino, C Garrino, P M Comoglio
JournalCancer research (Cancer Res) Vol. 49 Issue 6 Pg. 1415-21 (Mar 15 1989) ISSN: 0008-5472 [Print] United States
PMID2466553 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Antibodies, Monoclonal
  • Antigens, Neoplasm
  • Antigens, Tumor-Associated, Carbohydrate
  • Epitopes
  • Glycolipids
  • Mucins
Topics
  • Animals
  • Antibodies, Monoclonal
  • Antigens, Neoplasm (immunology, isolation & purification)
  • Antigens, Tumor-Associated, Carbohydrate (immunology)
  • Epitopes (analysis)
  • Glycolipids (immunology)
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Mucins (immunology)

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